Figure Legends
Figure 1: The relative abundance of WHY1 transcripts in the
base, middle and tip sections of the first leaf of 7-day old wild type
seedlings estimated by qRT-PCR (A) and Western blot (B). Data in panel A
are presented as means ± SE (n = 3). Different letters represent
statistical differences assessed by One-way ANOVA followed by
Tukey’s post hoc test.
Figure 2: Distribution of WHY1 in barley leaf determined by
immunogold labelling. Leaf sections were prepared for immunogold
labelling and the presence of WHY1 detected by ployclonol antibody as
described. Images are representative of plastids (p) in the leaf base
(A); chloroplasts (c) in the leaf tip (B); nucleus with nucleolus (nu)
in the leaf base (m, mitochondrion) (C) and a detail of the nucleus in
the leaf base (D). Example gold particles are indicated by white arrows.
Scale bars representing 0.5 (A, B, D) or 1 µm (C) are provided in
individual panels.
Figure 3: Cluster analysis comparison of abundance of
transcripts that differ significantly in the base, middle and tip
regions of wild type 7-day old barley leaves. Relative transcript
abundance is represented according to the legend shown. Transcripts were
grouped in clusters (A-E) as indicated and are ordered as listed in
table S1.
Figure 4: Cluster analysis comparison of abundance of key
transcripts that show a developmental pattern of expression in the base,
middle and tip regions of wild type, W1-1 and W1-7 barley leaves.
Relative transcript abundance is represented according to the legend
shown. Transcripts mentioned in the text are indicated and other
transcripts are ordered as indicated in table S3.
Figure 5: Cluster analysis comparison of abundance of
transcripts associated with light signalling and plastid development
exhibiting significant differences in abundance in base, middle and tip
regions of wild type 7 day old barley leaves. Relative transcript
abundance is represented according to the legend shown. Gene accession
numbers are indicated to the right of each row and further details are
provided in table S4.
Figure 6: Cluster analysis comparison of abundance of
transcripts associated with plastid biogenesis and development
exhibiting significant differences in abundance in the base of wild
type, W1-1 and W1-7 7 day old barley leaves. Relative transcript
abundance is represented according to the legend shown. Transcripts are
ordered as indicated in Table S6.
Figure 7: Polar and non-polar metabolites exhibiting
significant differences in abundance in base, middle and tip regions of
wild type 7 day old barley leaves. Abundance is shown relative to basal
regions for each compound where bars represent mean and lines SE (n=4).
■, base; □, middle; ■, tip. C14:0, tetradecanoic acid; C17:0,
heptacanoic acid; C22 alc, docosanol; C24 alc, tetracosanol.
Figure 8: Relative abundance of transcripts associated with
primary metabolism (A) and primary metabolites (B) exhibiting
significant differences in abundance in the basal region of 7-day old
wild type and WHY1 knockdown barley leaves.
Figure 9: Western blots of selected chloroplast proteins in the
base, middle and tip sections of the first leaves of wild type (WT) and
W1-1 and W1-7 of seedlings 7 days after germination. Proteins detected
are chlorophyll a/b-binding proteins: LHCB1 and LHCB2, the small subunit
of RUBISCO (RBCS), chloroplast ribosomal protein S1 (RPS1), , the large
subunit of RUBISCO (RBCL), the photosystem II protein (D1) and WHIRLY1
(WHY1).
Figure S1: Representative transmission electron micrographs of
nucleus in base of WT barley leaves indicating the association of WHY1
protein (red arrows) with electron dense chromatin (Ch).
Figure S2: A comparisons of leaf chlorophyll content in the
base, middle and tip sections of the first leaves of 7-day old wild type
(WT), W1-1 and W1-7 seedlings. Data are presented as mean ± SE (n = 6).
Different letters represent statistical differences assessed by one-way
ANOVA followed by Tukey’s post hoc test.